河南华溪蟹nanos1、nanos2、piwi基因部分序列的克隆及表达特征

Cloning of the partial sequences of nanos1,nanos2 and piwi genes and analysing of their expression patterns in the Sinopotamon henanense

为获取河南华溪蟹(Sinopotamon henanense)nanos1、nanos2和piwi基因的表达特征,在前期河南华溪蟹卵巢组织转录组测序的基础上,对生殖相关基因nanos1、nanos2和piwi进行了克隆和分析,首次获得了河南华溪蟹nanos1、nanos2和piwi基因的部分cDNA序列。利用半定量PCR获取了河南华溪蟹nanos1、nanos2和piwi基因的组织表达谱特征和在卵巢发育过程中的表达变化规律。研究结果显示,nanos1基因在卵巢、精巢、鳃、心和肌肉中均有表达;nanos2基因在卵巢、精巢、副性腺、心脏、肠和肌肉中有较高表达,且卵巢中表达量高于精巢中表达量;piwi基因特异表达于卵巢和精巢。卵巢发育中3个基因的表达时序研究显示,nanos1基因从卵巢发育的增殖期、小生长期、大生长期到成熟期表达量逐渐升高,与之相反,nanos2和piwi基因的表达呈现相反的趋势。这些研究结果将为河南华溪蟹nanos1、nanos2和piwi基因的功能分析奠定基础。

The nanos1,nanos2 and piwi genes in Sinopotamon henanense were cloned and analyzed,by the results of the ovary transcriptome sequences to obtain the expression characteristics of this species.The partial cDNA sequences of nanos1,nanos2 and piwi genes were obtained.The tissue expression pattern of nanos1,nanos2 and piwi genes in S.henanense were revealed by semi-quantitative PCR.The results showed that nanos1 gene was expressed in ovary,testis,sputum,heart and muscle.The nanos2 gene was highly expressed in ovary,testis,accessory gonad,heart,intestine and muscle,and especially the expression level in the ovary was higher than that in the testis.The piwi gene was expressed specifically in ovary and testis.Furthermore,the expression characterisations of these three genes during the ovary development were detected,which showed a gradually increasing expressing level trend of nanos1 gene in the ovary from the proliferative phase,small growth phase,large growth phase to mature phase.Comparatively,the expression trends of nanos2 and piwi genes showed the opposite patterns.Our studies provide the bases for the research on the functions of nanos1,nanos2,piwi genes in S.henanense.